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Agar

琼脂

Company: Sigma-Aldrich
Catalog#: A1296
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RNA Stability Measurements Using RT-qPCR in Arabidopsis Seedlings
Author:
Date:
2020-07-20
[Abstract]  Steady-state mRNA levels are determined by both the rates of transcription and degradation. Regulation of mRNA stability and/or degradation are key factors that can significantly affect mRNA levels and its biological functions. mRNA stability can be measured indirectly after transcription inhibition. This protocol described a rapid and sensitive method of mRNA stability measurement through quantitative reverse transcription PCR (RT-qPCR) after inhibition of RNA transcription by cordycepin in Arabidopsis seedlings. [摘要]  [摘要] 稳态mRNA的水平取决于转录和降解的速率。mRNA稳定性和/或降解的调节是可以显着影响mRNA水平及其生物学功能的关键因素。mRNA的稳定性可以在转录抑制后间接测量。该协议描述了通过拟南芥幼苗中的虫草素抑制RNA转录后,通过定量逆转录PCR(RT-qPCR)进行mRNA稳定性测定的快速灵敏方法。

[背景] mRNA稳定性的调控是基因表达调控的关键控制点。mRNA的稳定性对基因表达,分子和细胞表型,以及最终对植物发育,防御和其他生物过程都具有深远的影响。多种方法,例如RNA印迹分析,原位杂交,可用于测量转录抑制后的mRNA稳定性。在此协议中,我们描述了一种快速灵敏的方法,通过虫草素抑制转录后,通过RT-qPCR测量mRNA的稳定性。虫草素或3'-脱氧腺苷是腺苷类似物(参见参考文献2 )。可以将3'-脱氧腺苷掺入RNA,并由于3'位置不存在羟基部分而抑制转录延伸和RNA合成(参见参考文献6 )。我们已经成功地使用了这种方便而灵敏的方法来测量拟南芥幼苗中几种低丰度mRNA 的稳定性,包括初级microRNA转录本(Jia 等,2017)。在这里,我们用详细的实验程序和数据分析方法介绍该协议。

Micropropagation of Prickly Pear by Axillary Shoot Proliferation
Author:
Date:
2018-07-05
[Abstract]  A protocol for the axillary bud proliferation of prickly pear (Opuntia; Cactaceae) is presented. This genus is widely used as a crop in the arid and semi-arid areas of the globe worldwide, providing numerous benefits for human and animal consumption. In vitro culture for axillary bud proliferation is of great use to obtain a large quantity of plants in a short period of time, with potential uses in production and for the preservation of endangered species of the Opuntia genus.

The optimal medium for Opuntia in vitro culture consists of Murashige and Skoog medium (MS) and L2 vitamins. To increase the yield of the axillary bud proliferation, we recommend the addition of plant growth regulators (PGRs). This work suggests a 15 d incubation in the ...
[摘要]  提出了刺梨( Opuntia; Cactaceae )腋芽增殖的方案。 该属广泛用作全球干旱和半干旱地区的作物,为人类和动物消费提供了许多益处。 腋芽增殖的体外培养对于在短时间内获得大量植物具有重要意义,可用于生产和保存罂粟的濒危物种 属。

仙人掌体外培养的最佳培养基由Murashige和Skoog培养基(MS)和L2维生素组成。 为了增加腋芽增殖的产量,我们建议添加植物生长调节剂(PGRs)。 这项工作表明在含有2.2mg / L苄基氨基嘌呤(BA)的培养基中培养15天,然后将外植体转移到不含PGR的培养基中。 我们还解释了如何使植物适应体外条件。

【背景】仙人掌(刺梨)属是仙人掌科家族的成员之一(Bravo-Hollis,1978)。虽然它原产于美洲,但目前生长在欧洲南部,非洲北部,澳大利亚,中东,西亚和世界其他地区的野生和商业种植园中(Ochoa和Barbera,1995; Kiesling和Metzing) ,2017)。仙人掌对干旱和半干旱环境以及生活在这些地区的人类群落产生了深远的影响,因为尽管在干旱地区生长,其生物量仍然很高(Acevedo et al。,1983)。在许多这样的领域, Opuntia 属被用于许多方面。 仙人掌的幼枝可以作为蔬菜食用;水果直接食用或加工成果冻,果汁或糖果(Barba et ...

A Microfluidic Device for Massively Parallel, Whole-lifespan Imaging of Single Fission Yeast Cells
Author:
Date:
2018-04-05
[Abstract]  Whole-lifespan single-cell analysis has greatly increased our understanding of fundamental cellular processes such as cellular aging. To observe individual cells across their entire lifespan, all progeny must be removed from the growth medium, typically via manual microdissection. However, manual microdissection is laborious, low-throughput, and incompatible with fluorescence microscopy. Here, we describe assembly and operation of the multiplexed-Fission Yeast Lifespan Microdissector (multFYLM), a high-throughput microfluidic device for rapidly acquiring single-cell whole-lifespan imaging. multFYLM captures approximately one thousand rod-shaped fission yeast cells from up to six different genetic backgrounds or treatment regimens. The immobilized cells are fluorescently imaged for over a ... [摘要]  整个寿命的单细胞分析极大地增加了我们对细胞老化等基本细胞过程的理解。为了观察整个寿命期间的个体细胞,必须从生长培养基中移除所有后代,通常通过手动显微切割。然而,手动显微切割费力,低通量,并且与荧光显微镜不兼容。在这里,我们描述了多路复用裂变酵母寿命显微解剖器(multFYLM)的组装和操作,这是一种用于快速获取单细胞全寿命成像的高通量微流体装置。 multFYLM从多达六种不同的遗传背景或治疗方案中捕获约一千个杆状裂殖酵母细胞。将固定的细胞荧光成像超过一周,而将子代细胞从装置中取出。得到的数据集产生记录每个细胞复制寿命的高分辨率多通道图像。我们预计multFYLM将广泛适用于裂殖酵母(Schizosaccharomyces pombe)和其他对称分裂的单细胞生物的单细胞整个寿命研究。

【背景】细胞衰老导致细胞功能的累积下降,最终导致死亡。大多数关于细胞衰老的研究侧重于模型单细胞生物的复制寿命,例如出芽酵母酿酒酵母(Nyström和Liu,2014; Wasko和Kaeberlein,2014; Wierman和Smith,2014; Ruetenik和Barrientos ,2015)。细胞的复制寿命(RLS)被定义为母细胞在其生命过程中产生的女儿的数量(Henderson和Gottschling,2008; Sutphin等人,2014)。 ...

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